DNA chip and reactive solid carrier

Abstract

A reactive solid carrier favorably employable for manufacturing DNA chip is composed of a solid carrier and a plurality of vinylsulfonyl groups or their reactive precursors each of which is fixed onto a surface of the solid carrier by covalent bonding via a linking group, and a method for producing a DNA chip is performed by bringing the reactive solid carrier into contact with nucleotide derivatives or their analogues having a reactive group which is reactive with the vinylsulfonyl group or reactive precursor fixed to the solid carrier.

1 - 38 . (canceled)  
     
     
         39 . A method for preparing a reactive solid carrier comprising a solid carrier and a plurality of vinylsulfonyl groups or their reactive precursors each of which is fixed onto a surface of the solid carrier by covalent bonding via a linking group wherein the vinylsulfonyl group or its reactive precursor linked to the linking group is represented by the following formula: 
         -L-SO 2 —X in which L represents a linking group, and X represents a group of —CR 1 ═CR 2 R 3  or —CHR 1 —CR 2 R 3 Y wherein each of R 1 , R 2  and R 3  independently is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; Y represents a halogen atom, —SO 2 R 11 , —OCOR 12 , —OSO 3 M, or a quaternary pyridinium group; R 11  is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; R 12  is an alkyl group having 1 to 6 carbon atoms, or a halogenated alkyl group having 1 to 6 carbon atoms; and M is a hydrogen atom, an alkali metal atom, or an ammonium group which comprises bringing a solid carrier having reactive groups on its surface into contact with disulfone compounds having the following formula:     X 1 —SO 2 -L 2 -SO 2 —X 2     in which L 2  represents a linking group, and each of X 1  and X2 represents a group of —CR 1 ═CR 2 R 3  or —CHR 1 —CR 2 R 3 Y wherein each of R 1 , R 2  and R 3  independently is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; Y represents a halogen atom, —SO 2 R 11 ,    —OCOR 12 , —OSO 3 M, or a quaternary pyridinium group; R 11  is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; R 12  is an alkyl group having 1 to 6 carbon atoms, or a halogenated alkyl group having 1 to 6 carbon atoms; and M is a hydrogen atom, an alkali metal atom, or an ammonium group.    
     
     
         40 . The method of  claim 39 , wherein the reactive groups are amino groups, mercapto groups or hydroxyl groups.  
     
     
         41 . A method for producing a solid carrier having nucleotide derivatives or their analogues fixed onto its surface, which comprises bringing a reactive solid carrier comprising a solid carrier and a plurality of vinylsulfonyl groups or their reactive precursors each of which is fixed onto a surface of the solid carrier by covalent bonding into contact with nucleotide derivatives or their analogues having a reactive group which is reactive with the vinylsulfonyl group or reactive precursor.  
     
     
         42 . The method of  claim 41 , wherein the nucleotide derivatives or the analogues are oligonucleotides, polynucleotides, or peptide nucleic acids.  
     
     
         43 . The method of  claim 41 , wherein the vinylsulfonyl group or its reactive precursor linked to the linking group is represented by the following formula: 
         -L-SO 2 —X in which L represents a linking group, and X represents a group of —CR 1 ═CR 2 R 3  or —CHR 1 —CR 2 R 3 Y wherein each of R 1 , R 2  and R 3  independently is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; Y represents a halogen atom, —SO 2 R 11 , —OCOR 12 , —OSO 3 M, or a quaternary pyridinium group; R 11  is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; R 12  is an alkyl group having 1 to 6 carbon atoms, or a halogenated alkyl group having 1 to 6 carbon atoms; and M is a hydrogen atom, an alkali metal atom, or an ammonium group.    
     
     
         44 . The method of  claim 43 , wherein X is a reactive group having the formula of —CR 1 ═CR 2 R 3 .  
     
     
         45 . A method comprising bringing a solid carrier having nucleotide derivatives or their analogues fixed onto its surface which is produced by the method of  claim 41  having nucleotide derivatives or their analogues fixed onto its surface into contact with oligonucleotides or polynucleotides which are complementary to the nucleotide derivatives or their analogues fixed onto the surface of the solid carrier in the presence of an aqueous solvent, so as to combine the complementary oligonucleotides or polynucleotides with the nucleotide derivatives or their analogues.  
     
     
         46 . The method of  claim 45  wherein the oligonucleotides or polynucleotides have a detectable label.  
     
     
         47 . A method for preparing a reactive solid carrier comprising a solid carrier and a plurality of vinylsulfonyl groups or their reactive precursors each of which is fixed onto a surface of the solid carrier by covalent bonding via a linking group wherein the vinylsulfonyl group or its reactive precursor linked to the linking group is represented by the following formula: 
         -L-SO 2 —X in which L represents a linking group, and X represents a group of —CR 1 ═CR 2 R 3  or —CHR 1 —CR 2 R 3 Y wherein each of R 1 , R 2  and R 3  independently is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; Y represents a halogen atom, —SO 2 R 11 , —OCOR 12 , —OSO 3 M, or a quaternary pyridinium group; R 11  is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; R 12  is an alkyl group having 1 to 6 carbon atoms, or a halogenated alkyl group having 1 to 6 carbon atoms; and M is a hydrogen atom, an alkali metal atom, or an ammonium group, which comprises bringing a solid carrier having reactive groups on its surface into contact with bifunctional compounds having the following formula:     X 1 —SO 2 -L 2 -X 3     in which L 2  represents a linking group, X 1  represents a group of —CR 1 ═CR 2 R 3  or —CHR 1 —CR 2 R 3 Y wherein each of R 1 , R 2  and R 3  independently is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; Y represents a halogen atom, —SO 2 R 11 , —OCOR 12 , —OSO 3 M, or a quaternary pyridinium group; R 11  is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; R 12  is an alkyl group having 1 to 6 carbon atoms, or a halogenated alkyl group having 1 to 6 carbon atoms; and M is a hydrogen atom, an alkali metal atom, or an ammonium group, and X 3  represents a maleimide group, a halogen atom, an isocyanate group, a thioisocyanate group, a succimidoxy group, an aldehyde group, or a carboxyl group.    
     
     
         48 . A method for producing a solid carrier having nucleotide derivatives or their analogues fixed onto its surface, which comprises bringing a reactive solid carrier comprising a solid carrier and a plurality of vinylsulfonyl groups or their reactive precursors each of which is fixed onto a surface of the solid carrier by covalent bonding via a hydrophilic polymer chain into contact with nucleotide derivatives or their analogues having a reactive group which is reactive with the vinylsulfonyl group or reactive precursor.  
     
     
         49 . The method of  claim 48 , wherein the nucleotide derivatives or the analogues are oligonucleotides, polynucleotides, or peptide nucleic acids.  
     
     
         50 . The method of  claim 48 , wherein the vinylsulfonyl group or its reactive precursor linked to the linking group is represented by the following formula: 
         -L-SO 2 —X in which L represents a hydrophilic polymer chain group, and X represents a group of —CR 1 ═CR 2 R 3  or —CHR 1 —CR 2 R 3 Y wherein each of R 1 , R 2  and R 3  independently is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; Y represents a halogen atom, —SO 2 R 11 , —OCOR 12 , —OSO 3 M, or a quaternary pyridinium group; R 11  is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; R 12  is an alkyl group having 1 to 6 carbon atoms, or a halogenated alkyl group having 1 to 6 carbon atoms; and M is a hydrogen atom, an alkali metal atom, or an ammonium group.    
     
     
         51 . The method of  claim 50 , wherein X is a reactive group having the formula of —CR 1 ═CR 2 R 3  in which each of R 1 , R 2  and R 3  has the meaning as defined in claim  27 .  
     
     
         52 . A method for producing a solid carrier having nucleotide derivatives or their analogues fixed onto its surface, which comprises bringing a reactive solid carrier comprising a solid carrier and a plurality of vinylsulfonyl groups or their reactive precursors a number of which are linked to a polymer chain which is fixed onto a surface of the solid carrier by covalent bonding at a number of sites which number is smaller than the number of vinylsulfonyl groups or their reactive precursors linked to the polymer chain into contact with nucleotide derivatives or their analogues having a reactive group which is reactive with the vinylsulfonyl group or reactive precursor.  
     
     
         53 . The method of  claim 52 , wherein the nucleotide derivatives or the analogues are oligonucleotides, polynucleotides, or peptide nucleic acids.  
     
     
         54 . The method of  claim 52 , wherein the vinylsulfonyl group or its reactive precursor linked to the polymer chain is represented by the following formula: 
         -L-(—SO 2 —X) k   in which L represents a polymer chain, X represents a group of —CR 1 ═CR 2 R 3  or —CHR 1 —CR 2 R 3 Y wherein each of R 1 , R 2  and R 3  independently is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; Y represents a halogen atom, —SO 2 R 11 , —OCOR 12 , —OSO 3 M, or a quaternary pyridinium group; R 11  is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, an aryl group having 6 to 20 carbon atoms, or an aralkyl group having 7 to 26 carbon atoms in which its alkyl group has 1 to 6 carbon atoms; R 12  is an alkyl group having 1 to 6 carbon atoms, or a halogenated alkyl group having 1 to 6 carbon atoms; and M is a hydrogen atom, an alkali metal atom, or an ammonium group, and k is an integer of 2 or more.    
     
     
         55 . The method of  claim 54 , wherein X is a reactive group having the formula of —CR 1 ═CR 2 R 3 .  
     
     
         56 . The method of  claim 39 , wherein the disulfone compound is 1,2-bis(vinylsulfonylamide)ethane.  
     
     
         57 . The method of  claim 39 , wherein the disulfone compound is 1,2-bis(vinylsulfonylamide)propane.  
     
     
         58 . The method of  claim 39  wherein the reactive groups are amino groups.